Inflammatory disease states are leading causes of death and disability and a major cost to healthcare in the United States. There is an epidemic of death and hospitalizations due to adverse effects from NSAIDs. TAI- LCx (Turmeric Anti-Inflammatory-Lipophilic Component X) is a novel anti-inflammatory discovered in previously unrecognized, pharmacologically-active constituents of turmeric (Curcuma longa L.), and has demonstrated high potential as a safer, more cost-effective treatment. TAI-LCx is distinct from curcumin and curcuminoids that have been heavily researched and well characterized. With a nuanced action on multiple inflammatory mediators while avoiding the COX pathway, TAI-LCx is expected to reduce clinical complications and reduce the staggering economic costs of existing treatments. The hypothesis of the proposed research is that one constituent (or a small group of small molecules) within TAI-LCx possesses specific, potent and clinically relevant anti-inflammatory (AI) activity. The purpose of the proposed research, therefore, is to test the hypothesis that improved and optimized methods for extraction of TAI-LCx can be identified, to purify to homogeneity the active constituent(s) and determine its molecular formula (thereby enabling in phase II efforts to complete structural determination), and to further characterize the AI action of TAI-LCx, to determine whether additional constituents of turmeric might be synergistically involved with TAI-LCx in producing the AI properties detected to date. The Specific Aims of the proposed project are as follows. Aim # 1: Optimize TAI-LCx extraction via supercritical fluid extraction (SFE) and characterize the active constituent(s). This will increase sample purity and commercial scalability, and lead to better characterization of the active principle, and is potentially a patentable innovation. Optimization of the SFE method followed by purification of the active constituents in TAI-LCx will enable scale-up and production of >50 g of TAI-LCx. The SFE method will be more specific, more efficient, more environmentally friendly, and more cost-effective than the solvent-based extraction method used originally (in the original patent) to generate active TAI-LCx. Aim # 2: Determine the constituents of TAI-LCx and identify the active compound(s). Further drug development will require detailed composition information. Samples from successful SFE extractions and subsequent purification containing TAI-LCx activity will be analyzed by high resolution mass spectrometry to determine the molecular formula of all components within the extracts and the (putative) structural identity of the active principle(s). Aim # 3: Determine the effect of TAI-LCx on other anti-inflammatory pathways, such as COX-1, NFkB, AP-1, and interleukins (IL-2, IL-6). This will demonstrate therapeutic significance and application, and provide a comparison with existing FDA approved therapies, as well as provide product differentiation and potentially add to the body of knowledge concerning inflammatory pathways.